1. Field of the Invention
The present invention relates to a cell culture assay, and more particularly, to a cell culture assay having a structure which can easily and massively produce cells and more accurately observe a process of culturing cells.
2. Discussion of Related Art
Cell migration means migration of living organisms or individual cells in response of various physical, chemical, and biological stimuli, and is deeply involved with various diseases and biological phenomena in a human body, such as AIDS, pathogenic and bacterial infections, arteriosclerosis, arthritis, periodontitis, psoriasis, cancer, multiple sclerosis, male infertility, asbestos poisoning, ozone poisoning, etc.
However, most conventional cell migration evaluating assays are used for two-dimensional cell migration, and thus could not help observing crawling of cells. Here, since cells actually exist in a three-dimensional space, the conventional assays frequently obtained results different from the real phenomenon.
While assays analyzing three-dimensional cell migration, tissue formation, morphological changes, cell proliferation, etc. have been developed abroad, they had limits to expose cells under specific conditions or simulate real three-dimensional migration occurring in living organisms, and thus did not effectively evaluate and digitize test results.
Here, when scaffolds constituting an extracellular matrix (ECM) are introduced into a microfluidic channel, all advantages of microfluidic technology may be maintained, and reactions of cells in evaluation environments such as cell migration and morphological changes, tissue formation, cell proliferation, etc. can be three-dimensionally simulated.
As a conventional art reflecting such advantages, International Patent No. WO2009/126524 is provided. According to the conventional art, three-dimensional simulation of angiogenesis is succeeded by introducing a collagen scaffold which is one of the ECMs, and results of studies on reactions between vascular endothelial cells and cancer cells, between liver cells and vascular endothelial cells, and between nerve cells.
Here, the microfluidic technology can provide microenvironments around a cell, allow real-time observation and accurate quantification of a reaction of cells, reduce an amount of cells or samples used in a test, and evaluate various test conditions.
In addition, a scaffold-integrating technique can three-dimensionally introduce cells, culture cells in various directions, such as inside and both sides of a scaffold. Therefore, various cells can be cultured at the same time, and thereby interaction of the cells and interaction between the cells and the scaffolds can be studied. For this reason, the scaffold-integrating technique can also be applied in development of medical materials. Moreover, effects of various materials including a nanomaterial, a drug, and a protein on cells may be three-dimensionally evaluated.
However, a microfluidic platform according to a conventional art needs a pole array having a size of several tens to several hundreds of micrometers to fix between scaffold channels. If there is no pole, scaffolds may be leaked into the channels, and therefore the conventional microfluidic platform cannot be applied. With reference, most scaffolds are present in a liquid type. They are solidified after being injected into a specific location in a channel, and therefore a pole is needed to block the injected scaffolds in the specific location before they are solidified.
However, the above-described microfluidic platform has a limit to an area in which cells are reacted due to the pole preventing leakage of the scaffolds, and is very difficult to massively produce cells. In addition, the pole is still seen during observation, which becomes a factor disturbing quantification, and the critical problem is that the cells are preferentially reacted with the pole rather than the scaffolds.